The major objectives of this research are: 1) to continue our studies to characterize components of specialized cisternae of endoplasmic reticulum which are lined with carbohydrates and which very likely serve as a transport system in neuronal processes; and 2) to continue a portion of our studies to relate the localization of macromolecules to the expression of different functions performed by neurons and glia. Macromolecules of nervous tissue membranes will be localized by: 1) the use of specific lectins which bind to different sugars and toxins which bind to specific receptors in combination with peroxidase labeling techniques to visualize sites of such binding; and 2) the use of antisera to specific brain macromolecules together with peroxidase conjugated antibodies to visualize tissue sites containing these macromolecules. These experiments will be performed on normal tissue and tissue in which axonal transport is blocked by a local cooling method. We will continue to characterize potential candidates for glycoproteins lining the specialized smooth membrane cisternae using lectin affinity chromatography, gel electrophoresis and immunocytochemical procedures. Antisera to those glycoproteins which do line the smooth membrane cisternae will be used in experiments to determine if transport is blocked when specific carbohydrates lining the cisternae are complexed with antibody protein. The possible involvement of carbohydrate lined cisternae in transport will be tested further by studying whether or not specific membrane probes will be taken up by neurons and become associated with the cisternae during transport and what factors influence both uptake into the cell and association of the probes with the cisternae. The role of coated vesicle proteins and other structural proteins such as Alpha-actinin in transport (particularly retrograde transport) will be studied using immunocyto-chemical methods to localize specialized regions of the synaptic terminal containing these proteins, and using antisera to the proteins to test their effect on blockage of cellular uptake of macromolecules and/or subsequent retrograde transport of the molecules.